Journal: Antimicrobial Agents and Chemotherapy

Article Title: First Penicillin-Binding Protein Occupancy Patterns of β-Lactams and β-Lactamase Inhibitors in Klebsiella pneumoniae

doi: 10.1128/AAC.00282-18

Figure Lengend Snippet: Principal component analysis of the log-transformed PBP IC50 data for our 13 tested compounds in two K. pneumoniae strains. The plot shows the clustering of compounds according to their positions on the first and second eigenvector. These two vectors explained 86.5% of the total variance. Compounds were grouped into two general clusters. The first cluster contained β-lactams that primarily targeted PBP3 but differed in their secondary targets. The second cluster was comprised of compounds that primarily targeted PBP2 or both PBPs 2 and 4. Of note, among compounds in the second cluster, the carbapenems, amoxicillin, and mecillinam had substantially lower IC50 for their primary targets than the β-lactamase inhibitors. Symbols for β-lactamase inhibitors are smaller due to their much higher IC50 for their primary PBP target relative to the primary PBP target IC50 of the β-lactams.

Article Snippet: Biapenem was purchased from TRC Canada (Toronto, Ontario, Canada); doripenem and avibactam were from MedChem Express (Monmouth Junction, NJ); imipenem and meropenem were from AK Scientific (Union City, CA); sulbactam was from TCI America (Portland, OR); tazobactam, amoxicillin, piperacillin, aztreonam, cefepime, and cefsulodin were from Chem-Impex International, Inc. (Wood Dale, IL); and mecillinam was from Molekula (Newcastle Upon Tyne, United Kingdom).

Techniques: Transformation Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Amlexanox and Forskolin Prevents Isoproterenol-Induced Cardiomyopathy by Subduing Cardiomyocyte Hypertrophy and Maladaptive Inflammatory Responses

doi: 10.3389/fcell.2021.719351

Figure Lengend Snippet: ALX and FSK combination preserves mice myocardial architecture during CCS. (A) Representative whole hearts from all experimental groups. (B) Representative Masson’s trichrome stained longitudinal section of whole hearts from all groups. (C) Representative Masson’s trichrome stained transverse section of whole hearts from all groups. (D–G) Representative Western blots and graphical presentations cleaved caspase 3, collagen I, and collagen III compared between Vhl and PCH mice, and ALX treatment, FSK treatment, and ALX and FSK combination treatment ( n = 4 hearts per treatment group). Western blots were performed in triplicates, and each protein band in the representative blot is an independent biological sample. (H,I) Representative microscopic images of Masson’s trichrome staining and collagen volume fraction (CVF) in the ventricular tissue sections from all groups. The plotted values are the means of the CVF from each mouse ( n = 6–8 fields of view per 5–7 sections per 8–16 hearts per group). (J,K) Representative microscopic images of wheat germ agglutinin (WGA) merged with DAPI staining and graphical presentation of measured cardiomyocyte diameters from ventricular tissue sectionings across all groups. The plotted values are the means of the cardiomyocyte sizes from each mouse ( n = 10–12 cells per five fields of view per five sections per six to seven hearts per group). Representative cardiomyocytes are marked in yellow boxes, and their zoomed-in (× 5) inserts to show hypertrophy are marked in red boxes. The Vhl mice group is representative of the Ctrl mice group in results illustrations due to similarity in their data. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 among the therapeutic groups; &&& p < 0.001 vs. Vhl; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. the therapeutic groups. CVF was estimated by dividing the collagen area with the total myocardial area and multiple by 100. Data are expressed as mean ± SEM. Data were analyzed using one-way ANOVA, followed by Tukey’s post hoc analysis.

Article Snippet: PCH preventive models were made by intraperitoneal injection of ALX (ab142825; Abcam) 2.5 mg/100 g/day and/or FSK (1099; Tocris Bioscience, United Kingdom) 0.5 mg/100 g/day.

Techniques: Staining, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: Amlexanox and Forskolin Prevents Isoproterenol-Induced Cardiomyopathy by Subduing Cardiomyocyte Hypertrophy and Maladaptive Inflammatory Responses

doi: 10.3389/fcell.2021.719351

Figure Lengend Snippet: ALX and FSK combination prevents proinflammatory responses aggravation in myocardia during CCS by preventing necrosis. (A) Graphical presentations of evaluated sera concentrations of troponin I from groups. The sera troponin I analysis was performed in triplicates ( n = 8 mice per treatment group). ∗ p < 0.05, ∗∗∗ p < 0.001, among the therapeutic groups; &&& p < 0.001 vs. ISO (PCH); ## p < 0.01, ### p < 0.001, vs. the therapeutic groups. (B,C) Representative CD68 IHC staining and graphical presentation of the extent of inflammatory cells infiltration into the myocardial of all groups. The Vhl mice group is representative of the Ctrl mice group in results illustrations due to similarity in their data ( n = 6–8 field of view per six to eight sections per six to seven hearts per group). (D–H) Graphical presentations of inflammatory cytokines (IL-1β, IL-6, TNFα, IL-10, and TGF-β) were evaluated by ELISA from all groups. The cytokines analysis was performed in triplicates ( n = 7–8 mice per treatment group). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, among the therapeutic groups; &&& p < 0.001 vs. Vhl; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. the therapeutic groups. Data are expressed as mean ± SEM. Data were analyzed using one-way ANOVA, followed by Tukey’s post hoc analysis.

Article Snippet: PCH preventive models were made by intraperitoneal injection of ALX (ab142825; Abcam) 2.5 mg/100 g/day and/or FSK (1099; Tocris Bioscience, United Kingdom) 0.5 mg/100 g/day.

Techniques: Immunohistochemistry, Enzyme-linked Immunosorbent Assay

Journal: Cell Death Discovery

Article Title: Knockdown of SUCLG2 inhibits glioblastoma proliferation and promotes apoptosis through LMNA acetylation and the mediation of H4K16la lactylation

doi: 10.1038/s41420-025-02856-4

Figure Lengend Snippet: A H4K16la and L-Lac expression patterns in shNC and shSUCLG2 cells of U251 and LN229 were determined using western blotting. B Cell precipitates were collected for cleavage under targets and tagmentation (CUT&Tag) assay to identify H4K16la binding peaks, and the heatmap shows the distribution of H4K16la peaks near the translation start site (TSS). C Comparison of shNC and shSUCLG2 peaks in LN229 cells. D Genomic distribution of H4K16la, which was annotated for its localisation (promoter, exon, intron, or intergenic) and quantified in relative and absolute amounts. E Volcano plots of upregulated versus downregulated genes after CUT&Tag assay for H4K16la binding. F The heat map shows genes downregulated around TSS in LN229 cells containing shNC and shSUCLG2 as detected by the CUT&Tag assay. G KEGG analysis of CUT&Tag-detected downregulated genes at H4K16la. H GO analysis of CUT&Tag-detected downregulated genes. I Volcano plot of differential gene expression in RNA-seq. J KEGG analysis of downregulated genes in shSUCLG2 versus shNC using RNA-seq. K GO analysis of shSUCLG2 versus shNC downregulated genes using RNA-seq. L Combined CUT&Tag and RNA-seq analyses were performed to identify potential downstream target intersections of H4K16la. M Integrated genomics viewer trajectory track of CUT&Tag data showing enrichment of H4K16la at BEST1, GRAMD4H, and MBD6 promoter regions. N LN229 and U251 cells were treated with shNC or shRNA, and DNA fragments were immunoprecipitated with H4K16la antibody and analysed by qPCR. O Representative western blotting images show the quantification of BEST1, MBD6, and GRAMD4 protein levels. P Western blotting of IL-6 and IL-8 in shNC or shRNA lines of LN229 and U251. * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant.

Article Snippet: The following antibodies were used: SUCLG2 (A8976, 1:1 200, ABclonal, Wobum, MA), Bax (60267-1-Ig, 1:1000, Proteintech, Rosemont, IL), PCNA (HRP-60097, 1:1 000, Proteintech), Caspase-3 (68773-1-Ig, 1:1000, Proteintech), Bcl-2 (12789-1-AP, 1:1000, Proteintech), β-actin (11313-2-AP, 1:1 500, Proteintech), Cyclin D1 (60186-1-Ig, 1:1500, Proteintech), DLAT (ab172617, 1:1000, Abcam, Cambridge, UK), LMNA (ab172617, 1:1000, Abcam), L-Lac (PTM-1401RM, 1:1000, Jingjie Bio, Nanjing, China), D-Lac (PTM-1429RM, 1:1000, Jingjie Bio), HIF-1α (H1alpha67, 1:1000, Abcam), H4K16la (PTM-122, 1:1000, Jingjie Bio), H4 (PTM-1015RM, Jingjie Bio), Total oxidative phosphorylation Rodent Antibody Cocktail (ab110413, 1:1000, Abcam), BEST1 (1:1000, ab259836, Abcam); GRAMD4 (1:1000, 24299-1-AP9, Proteintech), MBD6 (1:1000, ab204403, Abcam); MFN1 (A21293, 1:1200, ABclonal); MFN2 (A19678, 1:1200, ABclonal); DR1 (A13298,1:1200, ABclonal); IL-6 (A26791, 1:1000, ABclonal); IL-8 (RP00052, 1:1000, ABclonal); anti-rabbit IgG (H + L) (ab205718, 1:5000, Abcam), and anti-mouse IgG (H + L) (ab205719, 1:5000, Proteintech).

Techniques: Expressing, Western Blot, Binding Assay, Comparison, Gene Expression, RNA Sequencing, shRNA, Immunoprecipitation